Obesity is an increasing health problem in the US, as it limits mobility and is an independent contributing factor to the development of diabetes and heart disease. Therefore, there is likely to be an ever increasing demand for Pharmaceuticals that may modulate the differentiation and development of adipocytes, and the lipid-associated metabolism pathways. The goal of our project (a phase 1, SBIR) is to develop an image-based high throughput, high content, screen to test the effects of candidate Pharmaceuticals on the differentiation and metabolic state of cultured human adipocytes. The project will be a collaborative effort between Vala Sciences Inc. (La Jolla, CA), which will develop the assay techniques, and Zen-Bio, Inc., (Research Triangle Park, NC), which will culture the cells and assist with the experiments and data interpretation. The Specific Aims of the project are: Specific Aim #1: To develop automated microscopy techniques and computer algorithms to accurately quantify the number, size, and distribution of lipid droplets in maturing human adipocytes. Specific Aim #2: To develop microscopy techniques for quantifying the expression levels and intracellular location of lipid-droplet associated proteins such as perilipin, adipophilin, TIP47, and hormone sensitive lipase, which are involved in fat droplet formation and metabolism. By the end of the Phase 1, we will have developed a "kit"-format product, which will include primary human preadipocytes, staining reagents to visualize the lipids, and software, which will allow potential customers to screen candidate Pharmaceuticals for their ability to modulate lipid droplet formation; we will also have made significant progress towards quantifying the expression levels and intracellular location of certain of the lipid droplet associated proteins. Phase 2 will likely involve further development of the software and staining techniques (particularly with regard to the proteins) and expansion of the product line to include additional cell types and other proteins of interest. [unreadable] [unreadable]